Authors
Tony Hazbun, Fulai Jin, and Jing Huang
Summary
The genome wide two-hybrid array is a spatially ordered set of yeast strains, each expressing a different protein fused to the Gal4 activation domain (AD)1, 2. A mating strategy is used where a strain expressing a protein fused to the Gal4 DNA binding domain (BD) is mated to each array strain. The strain expressing the Gal4 BD fusion protein (in PJ69-4α) is termed the “bait” and the array strains (PJ69-4A) expressing the Gal4 AD fusion protein are termed the “prey”. The protocol used here will reduce the screening burden involved in using a two-hybrid array. Additional benefit is that strains are represented multiple times hence allowing the reproducible positives to be easily identified3. The smart pooling procedure can be applied to other types of genomic arrays if they have similar screening and assay criteria as the two-hybrid array4.Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.Advertisement
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